Hepatic glycogen is formed by direct and indirect pathways whose activities reflect altered nutrition or disease. Direct/indirect pathway measurements often involve test meals where ∼ 10% of carbohydrate is galactose, but its effects on direct/indirect pathway estimates are unknown. Therefore, direct/indirect pathway contributions in 24-h fasted rats given 2g/kg 100% glucose (GLU, n=6) or 90% glucose-10% galactose (GLU+GAL, n=6) were measured by [U-(13)C]glucose dilution and by position-5/position-2 glycogen enrichment (H5/H2) from 2H(2)O. For GLU+GAL, galactose glycogenesis was independently measured with [1-(13)C]galactose. Glycogenesis was equivalent in both groups but for GLU+GAL, 23 ± 4% of glycogen was derived from galactose. [U-(13)C]glucose reported a 30 ± 3% direct pathway contribution to glycogenesis for GLU but only 20 ± 3% for GLU+GAL (p=0.012 vs. GLU). H5/H2 yielded identical direct pathway estimates (32 ± 3% GLU, 29 ± 6% GLU+GAL). Thus, galactose glycogenesis was undetected by H5/H2 while [U-(13)C]glucose reported a reduced direct/indirect pathway ratio. With [1-(13)C]galactose also present, correct glycogenic source contributions were obtained.
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